• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Measurement of protein translocation by SecA ATPase

Research Project

Project/Area Number 21K19226
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionNara Institute of Science and Technology

Principal Investigator

Tsukazaki Tomoya  奈良先端科学技術大学院大学, 先端科学技術研究科, 教授 (80436716)

Project Period (FY) 2021-07-09 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2022: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2021: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords膜タンパク質 / ナノディスク / 高速AFM / 膜蛋白質 / タンパク質
Outline of Research at the Start

細胞質で合成された新規のタンパク質の多くは生体膜を越えて輸送される。細菌では,SecYEG複合体とSecA ATPaseが複合体を形成し,タンパク質を膜透過させている。しかしながら,未だ基質タンパク質がどのように輸送されているのかその分子メカニズムは解明されておらず,反応最小単位での精密な動的観測が必要とされている。本研究では,原子間力顕微鏡(AFM)でタンパク質の膜透過を測定する。

Outline of Final Research Achievements

In eubacteria, the Sec translocon complex consisting of membrane proteins SecY, SecE, and SecG (SecYEG) functions as a passive channel for membrane protein translocation from the cytoplasm to the periplasm. A motor SecA ATPase for the protein translocation pushes pre-proteins into the SecYEG. The detailed dynamics of how the conformational change of SecA ATPase functions in concert with protein translocation is unclear. In this study, we attempted to visualize the structural dynamics of the SecYEG-SecA complex in protein translocation using high-speed atomic force microscopy. We were able to observe the binding of unfolded substrate proteins to the SecYAEG complex reconstituted into nanodiscs and the transport of the substrate proteins in real-time. In addition, nucleotide-dependent conformational changes of a domain of the SecA ATPase were visualized.

Academic Significance and Societal Importance of the Research Achievements

本研究は未だ可視化されていないSecタンパク質によるタンパク質の膜透過のダイナミクスを明らかとすべく研究を進めた。世界で初めてタンパク質輸送をリアルタイムで捉えることができた。今後は、この結果をまとめて原著論文として発表する。本解析は、機能解析が進んでいない他の膜タンパク質にも利用できるため、新しいことを発見した学術的意義だけでなく、膜タンパク質の解析法の一つとして技術的な貢献がある。

Report

(3 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • Research Products

    (13 results)

All 2022 2021 Other

All Int'l Joint Research (1 results) Journal Article (4 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 4 results,  Open Access: 4 results) Presentation (6 results) (of which Invited: 5 results) Remarks (2 results)

  • [Int'l Joint Research] 復旦大学(中国)

    • Related Report
      2022 Annual Research Report
  • [Journal Article] Crystal structure of the lipid flippase MurJ in a “squeezed” form distinct from its inward- and outward-facing forms2022

    • Author(s)
      Kohga Hidetaka、Mori Takaharu、Tanaka Yoshiki、Yoshikaie Kunihito、Taniguchi Katsuhide、Fujimoto Kei、Fritz Lisa、Schneider Tanja、Tsukazaki Tomoya
    • Journal Title

      Structure

      Volume: 30 Issue: 8 Pages: 1088-1097

    • DOI

      10.1016/j.str.2022.05.008

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Generation of stable microtubule superstructures by binding of peptide-fused tetrameric proteins to inside and outside2022

    • Author(s)
      Inaba Hiroshi、Sueki Yurina、Ichikawa Muneyoshi、Kabir Arif Md. Rashedul、Iwasaki Takashi、Shigematsu Hideki、Kakugo Akira、Sada Kazuki、Tsukazaki Tomoya、Matsuura Kazunori
    • Journal Title

      Science Advances

      Volume: 8 Issue: 36

    • DOI

      10.1126/sciadv.abq3817

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Inner membrane YfgM-PpiD heterodimer acts as a functional unit that associates with the SecY/E/G translocon and promotes protein translocation2022

    • Author(s)
      Miyazaki Ryoji、Ai Mengting、Tanaka Natsuko、Suzuki Takehiro、Dhomae Naoshi、Tsukazaki Tomoya、Akiyama Yoshinori、Mori Hiroyuki
    • Journal Title

      Journal of Biological Chemistry

      Volume: 298 Issue: 11 Pages: 102572-102572

    • DOI

      10.1016/j.jbc.2022.102572

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Crystal structures of a nicotine MATE transporter provide insight into its mechanism of substrate transport2021

    • Author(s)
      Tanaka Yoshiki、Iwaki Shigehiro、Sasaki Akira、Tsukazaki Tomoya
    • Journal Title

      FEBS Letters

      Volume: 595 Issue: 14 Pages: 1902-1913

    • DOI

      10.1002/1873-3468.14136

    • NAID

      120007166836

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Sec 複合体によるタンパク質膜透過の1ユニット動態観察2022

    • Author(s)
      塚崎智也、長池航、板家成良、春山隆充、菅野泰功、宮崎亮次、市川宗厳、内橋貴之
    • Organizer
      第22回日本蛋白質科学会年会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] タンパク質膜透過・輸送に関わるPpiD/YfgM複合体のin vivo光架橋解析2022

    • Author(s)
      宮崎亮次、鈴木健裕、堂前直、塚崎智也
    • Organizer
      第22回日本蛋白質科学会年会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] メガダルトン生命機能深化ダイナミクス2022

    • Author(s)
      塚崎智也
    • Organizer
      BioneX 生命科学の変革 公開シンポジウム 2022
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Secの動的探査など進捗状況報告2022

    • Author(s)
      塚崎智也
    • Organizer
      第3回構造生命科学研究会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] タンパク質の膜透過に関わるプロトン駆動型モーターSecDFの構造生命科学2022

    • Author(s)
      塚崎智也
    • Organizer
      第95回日本細菌学会総会
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] 高速AFMによるタンパク質膜透過装置Sec複合体の動的構造解析2021

    • Author(s)
      板家成良, 長池航, 春山隆充, 市川宗厳, 菅野泰功, 宮﨑 亮次, 内橋貴之, 塚崎智也
    • Organizer
      第17回21世紀大腸菌研究会
    • Related Report
      2021 Research-status Report
  • [Remarks] 塚崎智也の業績一覧

    • URL

      https://bsw3.naist.jp/tsukazaki/member_tsukazaki.html

    • Related Report
      2022 Annual Research Report
  • [Remarks] 奈良先端大・構造生命科学

    • URL

      https://bsw3.naist.jp/tsukazaki/

    • Related Report
      2021 Research-status Report

URL: 

Published: 2021-07-13   Modified: 2024-01-30  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi